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Excision of a selectable marker in transgenic rice (Oryza sativa L.) using a chemically regulated Cre/loxP system.

Removal of a selectable marker gene from genetically modified (GM) crops alleviates the risk of its release into the environment and hastens the public acceptance of GM crops. Here we report the production of marker-free transgenic rice by using a chemically regulated, Cre/loxP-mediated site-specific DNA recombination in a single transformation. Among 86 independent transgenic lines, ten were found to be marker-free in the T0 generation and an additional 17 lines segregated marker-free transgenic plants in the T1 generation. Molecular and genetic analyses indicated that the DNA recombination and excision in transgenic rice were precise and the marker-free recombinant T-DNA was stable and heritable.

Base Sequence↗

Biological control in greenhouse systems.

The controlled environment of greenhouses, the high value of the crops, and the limited number of registered fungicides offer a unique niche for the biological control of plant diseases. During the past ten years, over 80 biocontrol products have been marketed worldwide. A large percentage of these have been developed for greenhouse crops. Products to control soilborne pathogens such as Sclerotinia, Pythium, Rhizoctonia and Fusarium include Coniothyrium minitans, species of Gliocladium, Trichoderma, Streptomyces, and Bacillus, and nonpathogenic Fusarium. Products containing Trichoderma, Ampelomyces quisqualis, Bacillus, and Ulocladium are being developed to control the primary foliar diseases, Botrytis and powdery mildew. The development of Pseudomonas for the control of Pythium diseases in hydroponics and Pseudozyma flocculosa for the control of powdery mildew by two Canadian research programs is presented. In the future, biological control of diseases in greenhouses could predominate over chemical pesticides, in the same way that biological control of greenhouse insects predominates in the United Kingdom. The limitations in formulation, registration, and commercialization are discussed, along with suggested future research priorities.

Canada↗

Recycling of treated domestic effluent from an on-site wastewater treatment system for hydroponics.

An alternative method to conserve water and produce crops in arid regions is through hydroponics. Application of treated wastewater for hydroponics will help in stripping off nutrients from wastewater, maximising reuse through reduced evaporation losses, increasing control on quality of water and reducing risk of pathogen contamination. This study focuses on the efficiency of treated wastewater from an on-site aerobic wastewater treatment unit. The experiment aimed to investigate 1) nutrient reduction 2) microbial reduction and 3) growth rate of plants fed on wastewater compared to a commercial hydroponics medium. The study revealed that the chemical and microbial quality of wastewater after hydroponics was safe and satisfactory for irrigation and plant growth rate in wastewater hydroponics was similar to those grown in a commercial medium.

Conservation of Natural Resources↗

Farming in space: environmental and biophysical concerns.

The colonization of space will depend on our ability to routinely provide for the metabolic needs (oxygen, water, and food) of a crew with minimal re-supply from Earth. On Earth, these functions are facilitated by the cultivation of plant crops, thus it is important to develop plant-based food production systems to sustain the presence of mankind in space. Farming practices on earth have evolved for thousands of years to meet both the demands of an ever-increasing population and the availability of scarce resources, and now these practices must adapt to accommodate the effects of global warming. Similar challenges are expected when earth-based agricultural practices are adapted for space-based agriculture. A key variable in space is gravity; planets (e.g. Mars, 1/3 g) and moons (e.g. Earth's moon, 1/6 g) differ from spacecraft orbiting the Earth (e.g. Space stations) or orbital transfer vehicles that are subject to microgravity. The movement of heat, water vapor, CO2 and O2 between plant surfaces and their environment is also affected by gravity. In microgravity, these processes may also be affected by reduced mass transport and thicker boundary layers around plant organs caused by the absence of buoyancy dependent convective transport. Future space farmers will have to adapt their practices to accommodate microgravity, high and low extremes in ambient temperatures, reduced atmospheric pressures, atmospheres containing high volatile organic carbon contents, and elevated to super-elevated CO2 concentrations. Farming in space must also be carried out within power-, volume-, and mass-limited life support systems and must share resources with manned crews. Improved lighting and sensor technologies will have to be developed and tested for use in space. These developments should also help make crop production in terrestrial controlled environments (plant growth chambers and greenhouses) more efficient and, therefore, make these alternative agricultural systems more economically feasible food production systems.

Agriculture↗

Nitrogen in global animal production and management options for improving nitrogen use efficiency.

Animal production systems convert plant protein into animal protein. Depending on animal species, ration and management, between 5% and 45 % of the nitrogen (N) in plant protein is converted to and deposited in animal protein. The other 55%-95% is excreted via urine and feces, and can be used as nutrient source for plant (= often animal feed) production. The estimated global amount of N voided by animals ranges between 80 and 130 Tg N per year, and is as large as or larger than the global annual N fertilizer consumption. Cattle (60%), sheep (12%) and pigs (6%) have the largest share in animal manure N production. The conversion of plant N into animal N is on average more efficient in poultry and pork production than in dairy production, which is higher than in beef and sheep production. However, differences within a type of animal production system can be as large as differences between types of animal production systems, due to large effects of the genetic potential of animals, animal feed and management. The management of animals and animal feed, together with the genetic potential of the animals, are key factors to a high efficiency of conversion of plant protein into animal protein. The efficiency of the conversion of N from animal manure, following application to land, into plant protein ranges between 0 and 60%, while the estimated global mean is about 15%. The other 40%-100% is lost to the wider environment via NH3 volatilization, denitrification, leaching and run-off in pastures or during storage and/or following application of the animal manure to land. On a global scale, only 40%-50% of the amount of N voided is collected in barns, stables and paddocks, and only half of this amount is recycled to crop land. The N losses from animal manure collected in barns, stables and paddocks depend on the animal manure management system. Relative large losses occur in confined animal feeding operations, as these often lack the land base to utilize the N from animal manure effectively. Losses will be relatively low when all manure are collected rapidly in water-tight and covered basins, and when they are subsequently applied to the land in proper amounts and at the proper time, and using the proper method (low-emission techniques). There is opportunity for improving the N conversion in animal production systems by improving the genetic production potential of the herd, the composition of the animal feed, and the management of the animal manure. Coupling of crop and animal production systems, at least at a regional scale, is one way to high N use efficiency in the whole system. Clustering of confined animal production systems with other intensive agricultural production systems on the basis of concepts from industrial ecology with manure processing is another possible way to improve N use efficiency.

Animal Feed↗

Evaluation of berseem clover in diets of ruminants consuming corn crop residues.

Berseem clover hay was compared to alfalfa hay provided at 0, 25, and 50% of the diet DM in mixtures with corn crop residues to wether lambs. Berseem clover hay had lower (P < .05) concentrations of NDF, ADF, and CP than alfalfa hay. The digestibility of DM, DMI, and N balance did not differ (P > .05) between diets containing alfalfa hay or berseem clover hay. To evaluate stockpiled berseem clover as a supplement for grazed corn crop residues, berseem clover and oats were incorporated into a corn-corn-oat/berseem clover crop rotation for 3 yr in replicated 6.1-ha fields. Two cuttings of oat-berseem clover hay were harvested each summer before forage was stockpiled for winter grazing. After corn grain harvest, multiparous and primiparous crossbred cows in midgestation were allotted to each field at 1.01 ha/cow to strip-graze corn crop residues with or without stockpiled berseem clover or allocated to replicated drylots for 98 to 140 d. Each group was offered alfalfa-grass hay as large bales to maintain a mean body condition score of 5 on a 9-point scale. Mean rates of total and digestible OM disappearance from grazed and ungrazed field areas of berseem clover and corn crop residues did not differ over the 3 yr. In vitro organic matter disappearance (IVOMD) tended to decrease more rapidly (P = .13) and NDF and ADF concentrations increased more rapidly (P < .05) in berseem clover than in corn crop residues. Seasonal BW change did not differ (P > .05) between winter management systems in any year, and seasonal body condition score changes did not differ (P > .05) between cows grazing corn crop residues and berseem clover and those maintained in a drylot in yr 2 and 3. Cows grazing corn crop residues with or without berseem clover required less (P < .05) hay than those maintained in drylot. Although the effects of berseem clover hay supplementation on the intake and digestibility of corn crop residues do not differ from alfalfa hay, the nutritional value of stockpiled berseem clover decreases rapidly during winter, limiting its value as a standing supplement for corn crop residues in late winter.

Animals↗

[Dynamic analysis of land use and land coverage in mid-eastern China during 1988-2000].

Using remote sensing (RS) and geographic information system (GIS) technologies and based on the documentary and field sampling data, this paper constructed and analyzed the 1988 and 2000 landscape maps of mid-eastern China. The mapping area of mid-eastern China was about 2.87 x 10(6) km2. With the rapid increase of economy and population, the area of farmland and town in mid-eastern China increased 10.921 x 10(5) hm2 and 13.754 x 10(5) hm2, accounted for a 2.14% and 19.25% increase, respectively from 1988 to 2000. At the same time, the forest area decreased 17.83 x 10(4) hm2, declined by 1.12%, because of the increasing demand for lumber provision and reclamation. The meadow area in 2000 increased 19.287 x 10(5) hm2, ascending 7.75%, while the coverage and quality of grassland declined, owing to the deteriorating environment. The untilled land in 2000 dropped 34.31%, contrast with 1988 by the reason of a large number of lands being exploited.

China↗

Transgenic plant-derived pharmaceuticals - the practical approach?

Production of biopharmaceuticals in transgenic plants would involve the creation of a new industry. Those transgenic plants, including staple food crops, could provide many benefits to people all over the world. However, the new industry might require a strict regulation system. It is probable that such a strict system would not be acceptable to Japan or to most developing countries. Many countries should use non-food crops for production of biopharmaceuticals and take on more simple systems. The new industry must develop strategies for promoting the benefits of transgenic plant-derived biopharmaceuticals on both the domestic and worldwide scales.

Animals↗

Atmospheric dynamics and bioregenerative technologies in a soil-based ecological life support system: initial results from Biosphere 2.

Biosphere 2 is the first man-made, soil-based, bioregenerative life support system to be developed and tested. The utilization and amendment of local space resources, e.g. martian soil or lunar regolith, for agricultural and other purposes will be necessary if we are to minimize the requirement for Earth materials in the creation of long-term off-planet bases and habitations. Several of the roles soil plays in Biosphere 2 are 1) for air purification 2) as a key component in created wetland systems to recycle human and animal wastes and 3) as nutrient base for a sustainable agricultural cropping program. Initial results from the Biosphere 2 closure experiment are presented. These include the accelerated cycling rates due to small reservoir sizes, strong diurnal and seasonal fluxes in atmospheric CO2, an unexpected and continuing decline in atmospheric oxygen, overall maintenance of low levels of trace gases, recycling of waste waters through biological regeneration systems, and operation of an agriculture designed to provide diverse and nutritionally adequate diets for the crew members.

Agriculture↗

Precaution and Participatory Integrated Assessment of GM crops in Spain.

Qualitative Participatory Integrated Assessment (PIA) has been carried out to unveil the different accounts and uses of the precautionary principle and of the precautionary approach in the management of Bt commercial crops in Spain. In particular, two main interpretations have been identified: a case by case precaution (or caution) and a systemic precaution. Three scenarios on the plausible causes and consequences of commercial GM crops policy futures and the role of the precautionary principle and of precaution in them were also developed. Further research found that these scenarios could be linked to two broader worldviews about different plausible societies, models of agriculture, and of the role of ethics in the management of science and technology. It is argued that such worldviews, which go beyond the bounded rationality of scientific expertise on commercial GM crops, are used by policy makers in Spain, rather than or in conjunction with expert assessments, to make complex decisions in situations of large uncertainties and high stakes.

Bacillus thuringiensis↗

Quantification of plant stress using remote sensing observations and crop models: the case of nitrogen management.

Remote sensing techniques offer a unique solution for mapping stress and monitoring its time-course. This article reviews the main issues to be addressed for quantifying stress level from remote sensing observations, and to mitigate its impact on crop production by managing cultural practices. The case of nitrogen fertilization is used here as a paradigm. The derivation of canopy state variables such as the leaf area index (LAI) and chlorophyll content (C(ab)) is first addressed. It is demonstrated that the inversion of radiative transfer models leads to useful estimates of these variables. However, because of the ill-posed nature of the inverse problem, better accuracy is achieved when using prior information on the distribution of the variables and when multiplying LAI by C(ab) to get canopy level chlorophyll content. This variable, LAIxC(ab) is well suited for quantifying canopy level nitrogen content. It is used for nitrogen stress evaluation by comparison with a reference unstressed situation which is, however, not easy to get in practice. The combination of remote sensing observations with crop models provides an elegant solution for stress quantification through assimilation approaches. It fuses several sources of information within our knowledge of the processes involved and accounts for the environmental budget which can be integrated when making decisions about cultural practices. Conclusions are drawn on the issues related to the retrieval of canopy state variables from remote sensing data, to the link between these observables and crop models, and to the assimilation approaches. Avenues for further research are finally discussed along with the required observation system.

Biosensing Techniques↗

Effect of interactions on the nutrient status of a tropical soil treated with green manures and inorganic phosphate fertilizers.

Integrated nutrient management systems using plant residues and inorganic P fertilizers have high potential for increasing crop production and ensuring sustainability in the tropics, but their adoption requires in-depth understanding of nutrient dynamics in such systems. This was examined in a highly weathered tropical soil treated with green manures (GMs) and P fertilizers in two experiments conducted in the laboratory and glasshouse. The treatments were factorial combinations of the GMs (Calopogonium caeruleum, Gliricidia sepium, and Imperata cylindrica) and P fertilizers (phosphate rocks [PRs] from North Carolina, China, and Algeria, and triple superphosphate) replicated thrice. Olsen P, mineral N, pH, and exchangeable K, Ca, and Mg were monitored in a laboratory incubation study for 16 months. The change in soil P fractions and available P was also determined at the end of the study. Phosphorus available from the amendments was quantified at monthly intervals for 5 months by 33P-32P double isotopic labeling in the glasshouse using Setaria sphacelata as test crop. The GMs were labeled with 33P to determine their contribution to P taken up by Setaria, while that from the P fertilizers was indirectly measured by labeling the soil with 32P. The P fertilizers hardly changed Olsen P and exchangeable cations during 16 months of incubation. The legume GMs and legume GM+P did not change Olsen P, lowered exchangeable Ca, and increased exchangeable K about threefold (4.5 cmol[+]kg(-1) soil) in the first 4 months, even as large amounts of NH4-N accumulated (approximately 1000 mg kg soil(-1)) and soil pH increased to more than 6.5. Afterwards, Olsen P and exchangeable Ca and Mg increased (threefold) as NH4+-N and soil pH declined. The legume GMs also augmented reversibly sorbed P in Al-P and Fe-P fractions resulting in high residual effect in the soil, while fertilizer-P was irreversibly retained. The GMs increased PR-P utilization by 40 to over 80%, mobilized soil P, and markedly enhanced uptake of N, K, Ca, and Mg. Thus GMs+PRs is an appropriate combination for correcting nutrient deficiencies in tropical soils.

Agriculture↗

Elimination of marker genes from transgenic plants using MAT vector systems.

We have developed an efficient system (Multi-Auto-Transformation [MAT] vectors) for the removal of marker genes and to increase the regeneration frequency of transgenic crops without using antibiotic selection, reducing their possible environmental impact. The MAT vector system is designed to use the oncogenes (ipt, iaaM/H, rol) of Agrobacterium, which control the endogenous levels of plant hormones and the cell responses to plant growth regulators, to differentiate transgenic cells, and to select marker-free transgenic plants. The oncogenes are combined with the site-specific recombination system (R/RS). At transformation, the oncogenes regenerate transgenic plants and then are removed by the R/RS system to generate marker-free transgenic plants. The choice of a promoter for the oncogenes and the recombinase (R) gene, the state of plant materials and the tissue culture conditions greatly affect efficiency of both the regeneration of transgenic plants and the generation of marker-free plants. We have evaluated these conditions in several plant species to increase their generation efficiency. This chapter describes our transformation protocols using MAT vectors.

Cloning, Molecular↗

Cow weights and reproduction on native rangeland and native rangeland-complementary forage systems.

Replicated cow herds of 15 Angus x Hereford (AH) and 15 Brahman x Hereford (BH) F1 cows each were placed on native rangeland (NR) or native rangeland-complementary forage (NRCF) systems. The NR was 8.1 ha of sagebrush-mixed prairie per cow-calf unit and the NRCF used the same mixed prairie complemented by .4 ha of double-cropped winter wheat and summer annual forage replacing about 30% of the mixed prairie for each cow unit. Calving seasons, spring for NR and fall for NRCF, were selected to complement the respective forage systems. Cow weights, reproductive performance, and population demographics were measured through seven calf crops to investigate their relationship to forage systems and cow breed types. Seasonal average cow weights increased from 484 +/- 7 (SE) kg during calf crop 1 to a peak weight of 568 +/- 8 kg at calf crop 5. Average weight of AH cows was 514 +/- 3 kg, compared with 539 +/- 3 kg for the BH cows (P < .01). Angus x Hereford and BH cows were 38 and 44 kg heavier, respectively, on the NRCF system than on the NR system. The average percentage of reproduction for AH cows was 90 +/- 2%, compared with 87 +/- 2% for the BH cows (P < .13). Both breeds had approximately 87 (AH) to 89% (BH) reproductive rate on the NR system. In contrast, AH cows had 94 +/- 2% reproduction on the NRCF system, compared with 84 +/- 2% for the BH cows.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Johnson Space Center's Regenerative Life Support Systems Test Bed.

The Regenerative Life Support Systems (RLSS) Test Bed at NASA's Johnson Space Center is an atmospherically closed, controlled environment facility for human testing of regenerative life support systems using higher plants in conjunction with physicochemical life support systems. The facility supports NASA's Advanced Life Support (ALS) Program. The facility is comprised of two large scale plant growth chambers, each with approximately 11 m2 growing area. The root zone in each chamber is configurable for hydroponic or solid media plant culture systems. One of the two chambers, the Variable Pressure Growth Chamber (VPGC), is capable of operating at lower atmospheric pressures to evaluate a range of environments that may be used in a planetary surface habitat; the other chamber, the Ambient Pressure Growth Chamber (APGC) operates at ambient atmospheric pressure. The air lock of the VPGC is currently being outfitted for short duration (1 to 15 day) human habitation at ambient pressures. Testing with and without human subjects will focus on 1) integration of biological and physicochemical air and water revitalization systems; 2) effect of atmospheric pressure on system performance; 3) planetary resource utilization for ALS systems, in which solid substrates (simulated planetary soils or manufactured soils) are used in selected crop growth studies; 4) environmental microbiology and toxicology; 5) monitoring and control strategies; and 6) plant growth systems design. Included are descriptions of the overall design of the test facility, including discussions of the atmospheric conditioning, thermal control, lighting, and nutrient delivery systems.

Atmospheric Pressure↗

Strategies for farmers and policy makers to control nitrogen losses whilst maintaining crop production.

The nitrogen (N) cycle is essentially 'leaky'. The losses of small amounts of nitrate to waters and of ammonia and nitrous oxide to the atmosphere are a part of the global biogeochemical N cycle. However, intensive agricultural production, industry and vehicle use have more than doubled the amount of 'reactive' N in the environment, resulting in eutrophication, ecosystem change and health concerns. Research has identified agricultural practices that cause large losses of N and, in some cases, developed solutions. This paper discusses the problems of maintaining productivity while reducing N losses, compares conventional with low input (integrated) and organic farming systems, and discusses wider options. It also looks at the need to integrate studies on N with other environmental impacts, set in the context of the whole farm system, to provide truly sustainable agricultural systems.

Animals↗

Determination of abamectin in soil samples using high-performance liquid chromatography with tandem mass spectrometry.

Abamectin, which is comprised of a mixture of avermectins B1a and B1b, is a natural pesticide used as an anti-parasitic agent in livestock, ornamental, and agricultural crops, which can potentially be transported to aquatic systems. These compounds are highly toxic to both aquatic vertebrates and invertebrates at low concentrations in water. This investigation developed high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) techniques to support automated extraction by an accelerated solvent extraction (ASE) system and chromatographic techniques to measure residues of avermectins in complex soil samples. HPLC along with atmospheric pressure chemical ionization (APCI) MS/MS was used for separation and determination of avermectin isomers in soil samples. Average method recovery for abamectin by UV was 91%, while detection by MS/MS resulted in a 68% recovery for abamectin. Individual method recoveries by MS/MS were 53.6% for avermectin B1a and 36.8% for avermectin B1b. The use of tandem technology eliminated matrix interferences and resulted in an approximately eight-fold increase in sensitivity.

Anthelmintics↗

Photodegradation of pesticides on plant and soil surfaces.

Photodegradation is an abiotic process in the dissipation of pesticides where molecular excitation by absorption of light energy results in various organic reactions, or reactive oxygen species such as OH*, O3, and 1O2 specifically or nonspecifically oxidize the functional groups in a pesticide molecule. In the case of soil photolysis, the heterogeneity of soil together with soil properties varying with meteorological conditions makes photolytic processes difficult to understand. In contrast to solution photolysis, where light is attenuated by solid particles, both absorption and emission profiles of a pesticide are modified through interaction with soil components such as adsorption to clay minerals or solubilization to humic substances. Diffusion of a pesticide molecule results in heterogeneous concentration in soil, and either steric constraint or photoinduced generation of reactive species under the limited mobility sometimes modifies degradation mechanisms. Extensive investigations of meteorological effects on soil moisture and temperature as well as development of an elaborate testing chamber controlling these factors seems to provide better conditions for researchers to examine the photodegradation of pesticides on soil under conditions similar to the real environment. However, the mechanistic analysis of photodegradation has just begun, and there still remain many issues to be clarified. For example, how photoprocesses affect the electronic states of pesticide molecules on soil or how the reactive oxygen species are generated on soil via interaction with clay minerals and humic substances should be investigated in greater detail. From this standpoint, the application of diffuse reflectance spectroscopy and usage or development of various probes to trap intermediate species is highly desired. Furthermore, only limited information is yet available on the reactions of pesticides on soil with atmospheric chemical species. For photodegradation on plants, the importance of an emission spectrum of the light source near its surface was clarified. Most photochemical information comes from photolysis in organic solvents or on glass surfaces and/or plant metabolism studies. Epicuticular waxes may be approximated by long-chain hydrocarbons as a very viscous liquid or solid, but the existing form of pesticide molecules in waxes is still obscure. Either coexistence of formulation agents or steric constraint in the rigid medium would cause a change of molecular excitation, deactivation, and photodegradation mechanisms, which should be further investigated to understand the dissipation profiles of a pesticide in or on crops in the field. A thin-layer system with a coat of epicuticular waxes extracted from leaves or isolated cuticles has been utilized as a model, but its application has been very limited. There appear to be gaps in our knowledge about the surface chemistry and photochemistry of pesticides in both rigid media and plant metabolism. Photodegradation studies, for example, by using these models to eliminate contribution from metabolic conversion as much as possible, should be extensively conducted in conjunction with wax chemistry, with the controlling factors being clarified. As with soil surfaces, the effects of atmospheric oxidants should also be investigated. Based on this knowledge, new methods of kinetic analysis or a device simulating the fate of pesticides on these surfaces could be more rationally developed. Concerning soil photolysis, detailed mechanistic analysis of the mobility and fate of pesticides together with volatilization from soil surfaces has been initiated and its spatial distribution with time has been simulated with reasonable precision on a laboratory scale. Although mechanistic analyses have been conducted on penetration of pesticides through cuticular waxes, its combination with photodegradation to simulate the real environment is awaiting further investigation.

Humans↗